BIBioTechBIApplyingValidationA case stuStage 1 –TECHLOGICBioTechLogic, Inc.BIO TECHLOGICBIO ®TECHIOLOGICLogic, Inc.IO ®g Q...
Agenda• Process Backgrou• Risk Assessment• DOE• Parameter Evaluat• ConclusionsTECHLOGICBIO TECHLOGICBIO ®BioTechLogic, Inc...
Process BackThe scope of this case stupdata obtained from small-Chromatography runs perlimits in experiments planlimits in...
Process BackTECHLOGICBIO TECHLOGICBIO ®BioTechLogic, Inc.kgroundSlide 4Company Confidential
Risk AssessmOperating Parameter Normal OperatingUV at Start of AEX Fraction Collection > 0.50 AUUV at End of AEX Fraction ...
Risk AssessmThe parameters selectedpthe UV at start of fractioncollection, gradient slopeD d t i blDependent variables ana...
DOEPattern Exp. No.:UV at the staof fractioncollectionpcollection(AU)−−−− 1 0.40000 2 0.50000 3 0 50000 3 0.5−++− 4 0.4000...
Experimental• A summary of the varIon Exchange ChromOutputs step yield anOutputs, step yield anlisted in the following• Ru...
ExperimentalRun Resin Load GradientRunNumberResin Load(g/L)GradientSlope (%)1 15 42 20 83 20 84 15 125 20 85 20 86 15 47 2...
Statistical dat• For evaluation of statistion protein quality and qsoftware package was uSquares model for EffecSquares mo...
Statistical Model PStep Yieldp• The final model used foreffects and two factor int• There were two significai ifi t i t ti...
Main Effect Levera. UV at Start Collectionc. Gradient SlopeTECHLOGICBIO TECHLOGICBIO ®BioTechLogic, Inc.rage Plots for Ste...
Statistical Model PPurity by RP-HPLy y• The final model used forprimary effects (excludintwo-factor interactionstwo factor...
Two-Factor InteraPurity by RP-HPLy yStep Yield (%)Gradient Slope (%/CV)0UV @ End (%)a. UV at End Collection x Gradient Slo...
Confirmation• There is one area of threpresents process failp pyield, run 8. In order toacceptance criterion isoperated wi...
Design SpaceSpaceSpaceTECHLOGICBIO TECHLOGICBIO ®BioTechLogic, Inc.e vs Defined ControlBased on the models generated forh ...
Conclusions• The results of this DOdesign space from the• Based on our findingsh f th thwe chose for the threeappropriate ...
Final ParametNormal OpeOperating ParameterNormal Ope(NOUV at the beginning ofpooling, % from peak max Target: >p g pUV at ...
Thank YouBioTechLogic Inc serBioTechLogic Inc. serbiopharmaceutical indwith companies to mwith companies to mresource and ...
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Applying QbD to Biotech Process Validation

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In this session from the Institute of Validation Technology's Validation Week Europe, Kurtis Epp and John Kandl discuss how to implement QbD to all three stages of process validation.

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Applying QbD to Biotech Process Validation

  1. 1. BIBioTechBIApplyingValidationA case stuStage 1 –TECHLOGICBioTechLogic, Inc.BIO TECHLOGICBIO ®TECHIOLOGICLogic, Inc.IO ®g QbD to Biotech Processn :udy in applying QbD to– Process DesignIVT Validation Week28-30 March 201128 30 March 2011Kurtis Epp, John KandlBioTechLogic, Inc.
  2. 2. Agenda• Process Backgrou• Risk Assessment• DOE• Parameter Evaluat• ConclusionsTECHLOGICBIO TECHLOGICBIO ®BioTechLogic, Inc.undtionSlide 2Company Confidential
  3. 3. Process BackThe scope of this case stupdata obtained from small-Chromatography runs perlimits in experiments planlimits in experiments planDesign of Experiments (DTECHLOGICBIO TECHLOGICBIO ®BioTechLogic, Inc.kgroundudy is to provide and evaluatey p-scale Ion Exchangerformed within defined parameterned and executed according toned and executed according toDOE).Slide 3Company Confidential
  4. 4. Process BackTECHLOGICBIO TECHLOGICBIO ®BioTechLogic, Inc.kgroundSlide 4Company Confidential
  5. 5. Risk AssessmOperating Parameter Normal OperatingUV at Start of AEX Fraction Collection > 0.50 AUUV at End of AEX Fraction Collection < 20% of maximum pElution Gradient8% per CVElution Gradient(20 – 100% B oveEquilibration volume ≥ 5 CVLoad flow rate 100 – 120 L/h (TargeWash flow rate 100 – 120 L/h (TargeElution flow rate 100 – 120 L/h (TargePre-Equilibration flow rate 100 – 120 L/h (TargeF1 F3: 0 2Fraction VolumeF1 – F3: 0.2F4+: 0.1 CVWash volume ≥2 CVFraction Mixing Speed 65 rpmFraction Mixing Time 20 - 30 minPool Mixing Time10 – 15 minPool Mixing TimePool Mixing Speed100 rpmPre-Equilibration volume ≥ 4 CVWFI Rinse volume ≥ 3 CVWFI Rinse flow rate 100 – 120 L/h (TargeEquilibration flow rate 100 – 120 L/h (Targeq ( gIn-Process Control LimitsColumn Load 15 – 25 g / L rColumn Bed Height 30 ± 3 cmColumn Backpressure duringEquilibration, Load, Wash, and Elution< 3 barEffl t H t d f E ilib ti± 0.3 pH Units of EqEffluent pH at end of Equilibrationp qBuffer pHEffluent Cond. at end of Equilibration± 1 mS/cm Units of EBuffer ConducEffluent UV at the end of Equilibration ZeroFraction Pooling Criteria (RP-HPLC) ≥ 95% Main PTECHLOGICBIO TECHLOGICBIO ®BioTechLogic, Inc.mentg Range Potential Effect(s) of Failure SEV OCC DET RPNU Product loss, quality 9 4 2 72peak height Product loss, quality 9 4 2 72VInconsistent quality 8 2 4 64r 10 CV)Inconsistent quality 8 2 4 64Product loss, quality 9 2 1 18et: 110 L/h) Inconsistent quality 4 1 3 12et: 110 L/h) Inconsistent quality 4 1 3 12et: 110 L/h) Inconsistent quality 4 1 3 12et: 110 L/h) Longer equilibration 4 1 3 12CVCVVProduct loss, quality 9 1 1 9Inconsistent quality 5 1 1 5Inconsistent quality 4 1 1 4n Inconsistent quality 4 1 1 4nNon-homogeneity, inconsistentsampling/yield4 1 1 4sampling/yieldNon-homogeneity, inconsistentsampling/yield4 1 1 4Longer equilibration 3 1 1 3Longer equilibration 3 1 1 3et: 110 L/h) Longer equilibration 1 1 1 3et: 110 L/h) Product loss, quality 3 1 1 3) , q yresinmquilibrationqEquilibrationctivityPeakSlide 5Company Confidential
  6. 6. Risk AssessmThe parameters selectedpthe UV at start of fractioncollection, gradient slopeD d t i blDependent variables anaand purity by RP-HPLC.TestinInput parameterLowerUV at the beginning offraction collection (AU)0.40UV at the end of pooling(% from peak max)10Gradient slope (% per CV) 4.0Resin Load, (g/L resin) 15TECHLOGICBIO TECHLOGICBIO ®BioTechLogic, Inc.mentd for evaluation in this study wereyn collection, UV at end of fractione, and resin load.l d t i i ldalyzed were protein recovery yieldng limitsgOutput parametersUpper0.60Purity by RP-HPLC (≥ 95%)Step yield (≥ 75%)3012.025Slide 6Company Confidential
  7. 7. DOEPattern Exp. No.:UV at the staof fractioncollectionpcollection(AU)−−−− 1 0.40000 2 0.50000 3 0 50000 3 0.5−++− 4 0.40000 5 0.5++−− 6 0.67 0 6+−−+ 7 0.6++++ 8 0.6+−+− 9 0.6−−++ 10 0.40000 11 0.5−+−+ 12 0.4Note: It is important to randomizeindependence of your observationmistake by the operator.TECHLOGICBIO TECHLOGICBIO ®BioTechLogic, Inc.artnUV at the endof poolingGradientslopeResin Load(g/L)(% peak max) (% per CV)(g/L)10 4 1520 8 2020 8 2020 8 2030 12 1520 8 2030 4 1510 4 2510 4 2530 12 2510 12 1510 12 2520 8 2030 4 25e the run order to assure thens and reduce the chances of aSlide 7Company Confidential
  8. 8. Experimental• A summary of the varIon Exchange ChromOutputs step yield anOutputs, step yield anlisted in the following• Runs that did not meeare shaded to indicateTECHLOGICBIO TECHLOGICBIO ®BioTechLogic, Inc.Resultsriable parameters for each of theatography runs as well as thend eluate purity by RP HPLC arend eluate purity by RP-HPLC aretable.et the defined acceptance criteriape run failureSlide 8Company Confidential
  9. 9. ExperimentalRun Resin Load GradientRunNumberResin Load(g/L)GradientSlope (%)1 15 42 20 83 20 84 15 125 20 85 20 86 15 47 25 48 25 128 25 129 15 1210 25 1211 20 811 20 812 25 4TECHLOGICBIO TECHLOGICBIO ®BioTechLogic, Inc.ResultsUV @ Start% PeakStep Yield(%)RP-HPLCPurity (%)UV @ StartCollectionHeight EndCollection(%) Purity (%)Acceptance Criteria≥ 45% ≥ 95%0.4 10 53 950.5 20 56 970.5 20 50 970.4 30 59 960 5 20 51 970.5 20 51 970.6 30 53 970.6 10 52 960 6 30 40 970.6 30 40 970.6 10 50 980.4 10 45 970.5 20 51 970.5 20 51 970.4 30 60 97Slide 9Company Confidential
  10. 10. Statistical dat• For evaluation of statistion protein quality and qsoftware package was uSquares model for EffecSquares model for Effec• The model was run sepof yield and purity by RP• The fractional factorial minclusion of all single facinteractions as model efinteractions as model efresource and time constwas not possible.TECHLOGICBIO TECHLOGICBIO ®BioTechLogic, Inc.ta analysisically significant effects of factorsuantity, the JMP statisticalused applying the Standard Leastct Leveragect Leverage.arately for two output parametersP-HPLC.model was initially run with thector and some two-factorffects (Resolution IV) Due toffects (Resolution IV). Due totraints, a higher resolution studySlide 10Company Confidential
  11. 11. Statistical Model PStep Yieldp• The final model used foreffects and two factor int• There were two significai ifi t i t ti Tsignificant interaction. Tgraphically represented iplots and interaction conpTECHLOGICBIO TECHLOGICBIO ®BioTechLogic, Inc.Parameter Estimates forassessment all four primaryp yteractions listed above.nt main effects and oneh l ihese conclusions werein the main effects leveragetour plots.pSlide 11Company Confidential
  12. 12. Main Effect Levera. UV at Start Collectionc. Gradient SlopeTECHLOGICBIO TECHLOGICBIO ®BioTechLogic, Inc.rage Plots for Step Yieldb. UV at End Collectiond. Resin LoadSlide 12Company Confidential
  13. 13. Statistical Model PPurity by RP-HPLy y• The final model used forprimary effects (excludintwo-factor interactionstwo factor interactions.TECHLOGICBIO TECHLOGICBIO ®BioTechLogic, Inc.Parameter Estimates forCr assessment included all fourng resin load) as well as threeSlide 13Company Confidential
  14. 14. Two-Factor InteraPurity by RP-HPLy yStep Yield (%)Gradient Slope (%/CV)0UV @ End (%)a. UV at End Collection x Gradient SlopeTECHLOGICBIO TECHLOGICBIO ®BioTechLogic, Inc.action Contour Plots forCStep Yield (%)Resin Load (g/L resin)0UV @ Start (AU)b. UV at Start Collection x Gradient SlopeSlide 14Company Confidential
  15. 15. Confirmation• There is one area of threpresents process failp pyield, run 8. In order toacceptance criterion isoperated within its defioperated within its defimodified and two additaugment the initial des• We chose to tighten thas it was determined tosignificant primary effesignificant primary effeanalysis.• None of the experimenpfailed the acceptance cconfirms the predictedstudyTECHLOGICBIO TECHLOGICBIO ®BioTechLogic, Inc.study.of Resultshe tested design space thatlure space with regard to stepp g po ensure that the step yieldalways met when the process isned PARs the design space wasned PARs, the design space wastional runs were performed tosign.e gradient slope upper limit to 11%o be easy to control and the mostect for step yield in our initialect for step yield in our initialntal runs for the tightened limitsgcriterion for step yield. Thisdesign space from the initialSlide 15Company Confidential
  16. 16. Design SpaceSpaceSpaceTECHLOGICBIO TECHLOGICBIO ®BioTechLogic, Inc.e vs Defined ControlBased on the models generated forh f th d fi deach of the defined processoutputs, a three-dimensional plotwas generated to graphically showthe process control space withd h i llregard to the experimentallydefined design spaceSlide 16Company Confidential
  17. 17. Conclusions• The results of this DOdesign space from the• Based on our findingsh f th thwe chose for the threeappropriate with the ex• As a result we tighteneAs a result we tightene• The IPC limits for resinset appropriately.TECHLOGICBIO TECHLOGICBIO ®BioTechLogic, Inc.E study confirm the predictedy pe initial study.we determined that the IPC limitsi t te input parameters werexception of Gradient Slope.ed our control limitsed our control limits.n load were also determined to beSlide 17Company Confidential
  18. 18. Final ParametNormal OpeOperating ParameterNormal Ope(NOUV at the beginning ofpooling, % from peak max Target: >p g pUV at the end of pooling, %from peak max TargeGradient slope, % per CV TargeGradient slope, % per CV TargePerformance Parameter InResin Load, g/L* Note: The claimed experimentally confirmed range is slightly tighter than thsupport the higher end of the range.TECHLOGICBIO TECHLOGICBIO ®BioTechLogic, Inc.ter Limitsrating Range Experimentally Confirmedrating RangeOR)Experimentally ConfirmedRanges> 0.5 AU 0.4 – 0.6 AUt: 20% 10 – 30%et: 8% 4 – 11%*et: 8% 4 11%n-Process Acceptance Criterion15 – 25 g/Lhe design space in that it takes the worst-case data point toSlide 18Company Confidential
  19. 19. Thank YouBioTechLogic Inc serBioTechLogic Inc. serbiopharmaceutical indwith companies to mwith companies to mresource and commeTECHLOGICBIO TECHLOGICBIO ®BioTechLogic, Inc.rves therves thedustry by collaboratingeet their developmenteet their development,ercialization needs.www.biotechlogic.comwww.processvalidation.comSlide 19Company Confidential
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